Project Summary Hepatocellular carcinoma (HCC) is a major cause of cancer-related death worldwide including the United States due to late detection and limited effective therapeutic options. Sorafenib is the only FDA-approved drug currently available with limited efficacy for a small window of time. Designing effective therapeutic strategies to combat HCC is a major long-term focus of the current proposal. This proposal is based on our recent observations that revealed: (i) treatment of various HCC cells with the natural compound Berberine (BBR) reduces cell viability, (ii) BBR increases apoptosis in combination with proapoptotic TRAIL, (iii) TRAIL-BBR combination- induced apoptosis requires active AMP-activated protein Kinase (AMPK) signaling, (iv) BBR activates AMPK, (v) BBR antagonizes ?-catenin expression and inhibits ?-catenin/TCF- mediated downstream signaling. Based on these we hypothesize that BBR-induced AMPK activation represents a novel therapeutic strategy for treating HCC, which via inhibiting ?- catenin/TCF axis can antagonize HCC progression. In this pilot proposal, using biochemical, cell biological and in vivo approaches we plan on generating strong supporting data to validate our hypothesis so more elaborate studies with BBR can be designed in the future. Two specific aims are proposed: (Aim 1) Elucidate the role of AMPK in mediating BBR-induced antagonism of HCC and its potential crosstalk with ?-catenin and (Aim 2) Determine the efficacy of BBR and AMPK in antagonizing HCC progression in a mouse model of HCC. While Aim 1 studies are expected to provide a mechanistic insight towards the signaling pathway by which BBR antagonizes HCC and reveal a potential crosstalk between AMPK and Wnt/?-catenin axes, those in Aim 2 will provide important preclinical information on the in vivo efficacy of BBR- mediated AMPK activation on HCC. Successful outcome of the studies proposed will provide newer insights towards developing BBR-based mono or combination therapeutic strategies in the future for the targeting of HCC.